TNT nitroreductase from a Pseudomonas aeruginosa strain isolated from TNT-contaminated soil

B. T. Oh, G. Sarath, Patrick J Shea

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Microbial degradation and detoxification of TNT (2,4,6-trinitrotoluene) in soils ultimately depends on the presence of effective intracellular or extracellular enzymes. TNT was rapidly degraded by an enzyme obtained from a Pseudomonas aeruginosa strain isolated from TNT-contaminated soil. The nitroreductase enzyme was purified to apparent electrophoretic homogeneity by sequential chromatography on phenyl-sepharose, hydroxyapatite, and ion exchange resins. The TNT-nitroreductase catalyzed reduction of TNT to 4-hydroxylamino-4,6-dinitrotoluene (4HADNT) and 4-amino-2,6-dinitrotoluene (4ADNT). TNT reduction to 4ADNT in cell-free enzyme preparations corresponded to 2ADNT production by intact cells. The enzyme required NADH as a cosubstrate and the optimal pH for the reaction was approximately 6.5. The purified enzyme also exhibited NADH diaphorase activity and its denatured molecular weight was approximately 54 kDa.

Original languageEnglish (US)
Pages (from-to)875-881
Number of pages7
JournalSoil Biology and Biochemistry
Volume33
Issue number7-8
DOIs
StatePublished - Jun 26 2001

Fingerprint

trinitrotoluene
Nitroreductases
Trinitrotoluene
Pseudomonas aeruginosa
polluted soils
Soil
enzyme
Soils
Enzymes
enzymes
Dihydrolipoamide Dehydrogenase
Ion Exchange Resins
ion exchange resins
Detoxification
hydroxyapatite
NAD(P)H dehydrogenase (quinone)
Durapatite
detoxification
Chromatography
biodegradation

Keywords

  • Bacteria
  • Enzyme
  • Explosive
  • Reduction

ASJC Scopus subject areas

  • Microbiology
  • Soil Science

Cite this

TNT nitroreductase from a Pseudomonas aeruginosa strain isolated from TNT-contaminated soil. / Oh, B. T.; Sarath, G.; Shea, Patrick J.

In: Soil Biology and Biochemistry, Vol. 33, No. 7-8, 26.06.2001, p. 875-881.

Research output: Contribution to journalArticle

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AB - Microbial degradation and detoxification of TNT (2,4,6-trinitrotoluene) in soils ultimately depends on the presence of effective intracellular or extracellular enzymes. TNT was rapidly degraded by an enzyme obtained from a Pseudomonas aeruginosa strain isolated from TNT-contaminated soil. The nitroreductase enzyme was purified to apparent electrophoretic homogeneity by sequential chromatography on phenyl-sepharose, hydroxyapatite, and ion exchange resins. The TNT-nitroreductase catalyzed reduction of TNT to 4-hydroxylamino-4,6-dinitrotoluene (4HADNT) and 4-amino-2,6-dinitrotoluene (4ADNT). TNT reduction to 4ADNT in cell-free enzyme preparations corresponded to 2ADNT production by intact cells. The enzyme required NADH as a cosubstrate and the optimal pH for the reaction was approximately 6.5. The purified enzyme also exhibited NADH diaphorase activity and its denatured molecular weight was approximately 54 kDa.

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